Method of preparing pentonic acids from pentoses



Patented'Mer- 2,463,784

sm'rnon or raeraama PENTONIC scms raonr rau'rosas Lewis B. Lockwood, Peoria, 111., assignor to the United States of America as represented by the Secretary of Agriculture No Drawing. Application August 9, 1946,

' Serial No. 689,350

14 Claims. (01. 195-41) (Granted under the act March 3, 1883, as

amended April 30, 1928; 370 O. G. 757) 1 This application is made under the act or March 3, 1883, as mended by the act of April 30, 1928, and the invention herein described, it patented, may be manufactured and used by or 2 stirrers may be used as suitable vessels in which to carry out the process.

Considerable latitude is possible in the selection of the nutrients to be used in the conduct for the Government of the United States of 5 of this invention. Thus, the KHaPO4, MgSO4, America for governmental purposes without the and urea may be varied in quantity, or may be payment to me of any royalty thereon. omitted, and corn steep liquor, vegetable decoc- This invention relates to a method of prepartions, liver extract, or any of the wide variety of ing pentonic acids from the corresponding aldonutrient materials now marketed for the culture pentoses by fermentation with bacteria of the of micro-organisms, may be substituted for the genus Pseudomonas. There are eight possible dehydrated yeast extract. aldo-pentoses. They are d-arabinose, l-arabi- The following examples illustrate representanose, d-lyxose, l-lyxose, d-ribose, l-rigose, Ig-xyltive procedures used in practicing this invention. ose, and l-xylose. Of these, only -ara nose, l-arabinose, d-ribose, and d-xylose are of com- EXAMPLE I mon occurrence. This invention relates, more The non-saccharine constituents of all of the particularly, to the oxidation of these four penculture solutions were the same. They constituttoses to the corresponding pentonic acids, respeced urea, 02 8111-; K i i. gm: M S04HHz0, tively, d-arabonic acid, l-arabonic acid, d-ribonic 8 de ydrated y ast extract, 0.5 gm.; soyacid, and d-xylonic acid. 20 P 3 d p P9 100 ml. culture; and CaCOz Fermentative oxidation ofsugars has a numexcess of the quantity n ss ry to neutralize ber 'of advantages over chemical oxidation of the maximum quantity f p n nic acid which sugars in t costly reagenm are t required could theoretically be formed from the pentose. better yields of oxidized products may be obtained The culture solutions were aerated y u in under suitable conditions, and the microbial oxi- 5 air through them at the rate of 100 ml. air per 100 dation processes are more specific, resulting in h P minuteh temp rature was fewer side reactions leading t cgntaminating maintained at C'. d-Arabinose was added to products which interfere with recovery the basal culture solution, and 100 ml. portions The xidation of arabinose to arabonjc acid Of were inoculated with bacteria Of the species: was reported by Bertrand (Compt. Rend. Acad. Pseudomonas f P. grave len, sy r n ha. Sci" Paris, 127912943 139 who used acetic and P. vendrelli. Analyses of mashes, made aftacid b t i Bertrand c t; Rend l er seven days, gave the data contained in Table I. Sci., Paris, 127:124-127 (1898)), also reported TABLEI the oxidation of xylose to xylonic acid by acetic v acidbacteria. The use of bacteria of the genus grams of d'ambmose in 100 culture 801% Pseudomonas for pentose oxidation has not been previously reported in the literature. The use of Pseudomonas instead of acetic acid bacteria Grams Weightyleld ofiers the advantages that the process is of wider q-m ni fif applicability, since acetic acid bacteria do not 40 tgri dt tfif mi-100 oxidize some of the pentoses which are readily culture .53 3 oxidized by bacteria of the Pseudomonas genus, and since pure synthetic nutrients may be used, Psmdommmgfmgii 20 2M although the addition of cmde nitrogenous nuifigg wag -53 trients may be made in some cases where these 'Pseudomonaa marezuzajj jj 1 1 will not interfere with recovery. It is well known that acetic acid bacteria require the addition of EXAMPLE n expensive vitamins if purely synthetic mashes are t be used s additions are not required by l-Arabinose instead of d-arabinose was added bacteria of the genus pseudomonas t0 the basal Culture Solution described in In this invention the mashes may be aerated, P01130118 were inoculated with baceither at atmospheric pressure or at super-atmostime Of the Species PSeudOmOMs fl s, pheric pressure by any means known to those fmy ild n i p fi yn-m l skilled in the art. Rotating drums, or deep vesand P. vendrelli. Analyses made after six days sels equipped with disperser stones, agitators, or 05 gave the data presented in Table II.

TABLE II 4.0 grams of l-arabinose per 100 ml. culture solution Grams, Weight yield l-arabonic gxgg acid produced acid 100 per 100 ml. grgms culture l-arabinose Paeudomonas fluorescent 6 1. 38 28. 2 Pseudomonas fragii 25 l. 75 35. 7 Pseudomonas mildenbcrgii 21.. 1. 41 28. 8 Pseudomonas putida 13.. 2. 34 47. 8 Pacudomonas sunzantha 79.- 86 17.5 Pacudomoncs vendrelli 23 2. 22 45. 4

EXAMPLE III cl-Ribose, instead of d-arabinose, was added to the basal culture solution described in Example I. 100 ml. portions were inoculated with bacteria of of the species Pseudomonas fluorescens, P. graveolens, P. mildenbergii, P. ovalis, P.-put ijaciens, P. synzcantha, and P. uendrelli. Analyses made after nine days gave the data in Table III.

TABLE III 5.9 grams d-ribose per 100 ml. culture solution Grams d- Weight yield ribonic acid grams dproduced ribonic acidper 100 ml. per 100 grams culture d-ribose Pseudomonas fragii 25 1.08 18. 4 Pseudomoriaa mildenberaii 21.- 0. 58 9. 9 Paeudomimas ovalis 8 0. 41 7.0 Pseudo'mzmas putri/acicnc 76 0.81 14.8 Paeudomonac aynzantha 79. l. 69 28.8 Pseudo-menus cendrelli 23... 2. 89 49. 3 Pscudomonas fluorescent 6. 4.11 70.0

EXAIVIPLE IV d-Xylose, instead of d-arabinose, was added to the basal culture solution described in Example I. 100 ml. portions were inoculated with bacteria of the species Pseudomonas Jiuorescens, P. fragii, P. graveolens, P. mildenbergz'i, P. ovalis, and P.

Having. thus described my invention, I claim:

1. A process of making a pentonic acid comprising inoculating a nutrient medium containing an aldo-pentose with bacteria of the genus Pseudomonas, and aerating the medium, a neutralizing agent for the pentonic acid formed being present.

2. A process of making d-arabonic acid coniprising inoculating a nutrient medium containing d-arabinose with bacteria of the genus Pseudomonas, and aerating the medium, a neutralizing agent for the pentonic acid formed being present.

3. A process of making d-ribonic acid comprising inoculating a medium containing d-ribose with bacteria of the genus Pseudomonas, and aerating the medium, a neutralizing agent for the pentonlc acid formed being present.

.4. A process of making d-xylonic acid comprising inoculating a medium containing d-xylose with bacteria of the genus Pseudomonas, and aerating the medium, a neutralizing agent for the pentonic acid formed being present.

5. The process of claim 1 in which the bacteria is Pseudomonas fluorescens.

6. The process of claim 1 in which the bacteria is Pseudomonas mildenbergii.

'7. The process of claim 2 in which the bacteria is Pseudomonas jragii.

8. The process of claim 3 in which the bacteria is Pseudomonas fluorescens.

9. The process of claim 4 in which the bacteria is Pseudomonas fluorescens.

10. The process of claim 4 in which the bacteria is Pseudomonas mildenbergii.

11. The process of claim 1 in which the neutralizing agent is a calcium compound.

12. The process of claim 1 in which the neutralizing agent is calcium carbonate and the temperature is maintained at about 30 C., the calcium carbonate being in excess of the quantity necessary to neutralize the maximum theoretical yield of pentonic acid from the pentose.

13. A process of converting d-xylose to dxylonic acid in high yields, comprising inoculating a nutrient medium containing d-xylose with Pseudomonas fluorescens, and aerating the medium, the said medium containing urea, KH2PO4, MgSO dehydrated yeast extract, and calcium carbonate in excess of the quantity necessary to neutralize the maximum theoretical yield of d-xylonic acid from d-xylose.

14. A process of making a pentonic acid comprising inoculating a nutrient medium containing an aldo-pentose with bacteria of the genus Pseudomonas, and aerating the medium, the said medium containing urea, KHzPO4, MgSO4, dehydrated yeast extract, and calcium carbonate in excess of the quantity necessary to neutralize the maximum theoretical yield of pentonicacid from the pentose.

LEWIS B. LOCKWOOD.

REFERENCES CITED UNITED STATES PATENTS Name Date Grandel Aug. 22, 1944 OTHER REFERENCES Lockwood et al., J. Bact., July 1941, p e 54. Chemical Abstracts, vol. 34, 7321, '7 and 8 Number (1940), citing V. V. Pervozvankii-Microbiology (U. S. S. R.) 8, N0. 2, 149-159 (1939) and N0. 3-4, 339-52. 

